U.S. patent application number 15/097104 was filed with the patent office on 2017-10-12 for methods and compositions for reducing contamination on food contact surfaces.
The applicant listed for this patent is WTI, Inc.. Invention is credited to Kevon LEDGERWOOD, Wolfgang LUDWIG, Jasdeep K. SAINI.
Application Number | 20170290342 15/097104 |
Document ID | / |
Family ID | 59999139 |
Filed Date | 2017-10-12 |
United States Patent
Application |
20170290342 |
Kind Code |
A1 |
SAINI; Jasdeep K. ; et
al. |
October 12, 2017 |
METHODS AND COMPOSITIONS FOR REDUCING CONTAMINATION ON FOOD CONTACT
SURFACES
Abstract
Embodiments disclosed herein provide methods for decontaminating
food contact surfaces using a fine spray/mist that comprise an
antimicrobial composition comprising an organic acid and an organic
citrus extract. The spray application reduces the microbiological
populations on the surface of food contact surfaces, including
potential pathogens, thereby reducing cross-contamination and
improving food safety. The composition described herein can be use
to target a number of microbiological pathogens, including L.
monocytogenes, which is a known cross-contaminant in retail
environments such as deli counters and slicers due to its ability
to grow at refrigerated temperatures and moist environments making
it a known cause of post-processing contamination of ready-to-eat
food products.
Inventors: |
SAINI; Jasdeep K.; (Athens,
GA) ; LEDGERWOOD; Kevon; (Jefferson, GA) ;
LUDWIG; Wolfgang; (Jefferson, GA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
WTI, Inc. |
Jefferson |
GA |
US |
|
|
Family ID: |
59999139 |
Appl. No.: |
15/097104 |
Filed: |
April 12, 2016 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
Y02A 40/90 20180101;
A61L 2/18 20130101; A01N 65/36 20130101; A61L 2202/20 20130101;
A01N 37/02 20130101; A01N 65/36 20130101; A01N 37/02 20130101 |
International
Class: |
A01N 65/36 20060101
A01N065/36; A61L 2/18 20060101 A61L002/18; A01N 37/02 20060101
A01N037/02 |
Claims
1. An antimicrobial composition comprising an organic acid and an
organic citrus extract, wherein the organic acid is a buffered
vinegar at a concentration of approximately 90% to approximately
99.5% w/v, wherein the buffered vinegar is buffered to a pH of
6.1.+-.0.1, and wherein the organic citrus extract is derived from
an albedo layer of a citrus fruit, a flavedo layer of a citrus
fruit, or a combination thereof and is at a concentration of 0.1%
to 5% w/v.
2. (canceled)
3. The antimicrobial composition of claim 1, wherein the organic
acid has a pH of approximately 5.5 to approximately 7.3.
4. The antimicrobial composition of claim 3, wherein the organic
acid has a pH of approximately 5.9 to approximately 7.1.
5. The antimicrobial composition of claim 1, wherein the organic
acid is at a concentration of between approximately 95% to
approximately 99.5% w/v acetic acid, formic acid, propionic acid,
butyric acid, valeric acid, caproic acid, oxalic acid, lactic acid,
malic acid, citric acid, benzoic acid, carbonic acid, or a
combination thereof.
6. (canceled)
7. The antimicrobial composition of claim 2, wherein the vinegar
comprises acetic acid and/or a consumable salt equivalent
thereof.
8. The antimicrobial composition of claim 1, wherein the vinegar
comprises 0.5% to 8% acetic acid.
9. The antimicrobial composition of claim 8, wherein the vinegar
comprises 1% to 4% acetic acid.
10. The antimicrobial composition of claim 6, wherein the vinegar
is a corn vinegar, a sugar cane vinegar, a glacial acetic vinegar,
an apple cider vinegar, or a combination thereof.
11. The antimicrobial composition of claim 1, wherein the organic
citrus extract is at a concentration of 0.1% to 1% w/v.
12. The antimicrobial composition of claim 1, further comprising a
masker, an emulsifier, or a combination thereof.
13. The antimicrobial composition of claim 12, wherein the masker
is at a concentration of 0.05% to 0.2% w/v.
14. A method for reducing microbiological contamination on food
contact surfaces comprising applying an antimicrobial composition
comprising an organic acid and an organic citrus extract in an
effective amount to reduce or eliminate a microbial population on
the food contact surface.
15. The method of claim 14, wherein the antimicrobial composition
has a organic acid concentration of approximately 50% to 99.5% w/v
and an organic citric extract concentration of approximately 0.5%
to approximately 5% w/v.
16. The method of claim 15, wherein the organic acid has a pH of
approximately 5.5 to approximately 6.2.
17. The method of claim 14, wherein the organic acid is acetic
acid, formic acid, propionic acid, butyric acid, valeric acid,
caproic acid, oxalic acid, lactic acid, malic acid, citric acid,
benzoic acid, and carbonic acid
18. The method of claim 17, wherein the acetic acid is provided as
a vinegar.
19. The method of claim 18, wherein the vinegar comprises 0.5% to
8% acetic acid.
20. The method of claim 18, wherein the vinegar is a corn vinegar,
a sugar cane vinegar, a glacial acetic acid vinegar, an apple cider
vinegar, or a combination thereof.
21. The method of claim 14, wherein the antimicrobial composition
is applied as a fine spray or mist.
22. The method of claim 14, wherein the antimicrobial composition
is allowed to remain in contact with the food contact surface for
at least 30 seconds prior to food coming into contact with the food
contact surface.
23. The method of claim 22, wherein the food contact surface is a
wood surface, a plastic surface, a rubber surface, a glass surface,
or a stainless steel surface.
24. The method of claim 14, wherein the microbial population
comprises one or more bacterial species and/or one or more fungal
species.
25. The method of claim 24, wherein the bacterial species is a
psychrotroph, a coliform, a lactic acid bacterial species, or a
spore-forming bacterial species.
26. The method of claim 25, wherein the bacterial species is a
Pseudomonas species, a Micrococcus species, an Aerococcus species,
a Lactococcus species, Leuconostoc species, a Streptococcus
species, a Bacillus species, a Clostridium species, a Eubacterium
species, an Enterococcus species, or a Listeria species.
27. The method of claim 26, wherein the bacterial species is a
Listeria species.
28. The method of claim 27, wherein the bacterial species is L.
monocytogenes.
29. The method of claim 24, wherein the one or more fungal species
comprises one or more yeast.
30. The method of claim 29, wherein the yeast is a Geotrichum
species, a Saccharomyces species, a Hansenula species, a Candida
species, a Kluyveromyces species, a Debaryomyces species, a Pichia
species.
Description
TECHNICAL FIELD
[0001] The invention relates to method for reducing microbiological
contamination on food contact surfaces, such as deli-slicers and
meat counters, using a fine spray/mist method of applying
compositions comprising organic acids in combination with organic
citrus extracts.
BACKGROUND
[0002] Listeria monocytogenes is a pathogenic bacterium that causes
listeriosis and has been implicated in several outbreaks linked to
consumption of ready-to-eat (RTE) sliced deli meats. RTE meat
slicers especially in retail delis provide ideal conditions for
certain bacteria, like Listeria, to colonize and grow. Sliced RTE
meats can become contaminated with this pathogen during the slicing
process and may pose a serious public health concern. Hence,
effective interventions are needed to control this pathogen and
prevent cross-contaminations from deli slicers on to the RTE food
product being sliced. Accordingly, methods for controlling such
contamination that are effective and safe for use with equipment
used to prepare food products is needed.
SUMMARY
[0003] The embodiments disclosed herein are directed to
antimicrobial compositions comprising an organic acid and an
organic citrus extract and use of said compositions in methods for
reducing microbial contamination on food contact surfaces. The
organic acid may be acetic acid, formic acid, propionic acid,
butyric acid, valeric acid, caproic acid, oxalic acid, lactic acid,
malic acid, citric acid, benzoic acid, carbonic acid, or a
combination thereof. In one example embodiment, the organic acid is
acetic acid. The organic citrus extract is derived from a blend of
two or more citrus fruit extracts. In certain example embodiments,
the organic citrus extract is a blend of two or more citrus fruit
extracts, excluding grapefruit derived extracts. Food contact
surfaces that may be treated with the antimicrobial composition
include, but are not limited to, stainless steel surfaces, glass
surfaces, rubber or wood surfaces. The antimicrobial compositions
may be used to reduce microbiological contamination caused by a
number of bacteria and fungal species, including Listeria species.
In certain example embodiments, the antimicrobial compositions are
applied to the food contact surfaces as a fine mist or spray.
[0004] In another aspect, the embodiments disclosed herein are
directed to methods for reducing microbiological contamination on
food contact surfaces comprising applying the antimicrobial
compositions described herein in an effective amount to reduce or
eliminate one or more microbial populations on a food contact
surface. The antimicrobial composition may be applied as a fine
spray or mist. The food contact surface may be a wood surface, a
plastic surface, a rubber surface, a glass surface, or a stainless
steel surface. Microbial populations that may be reduced or
eliminated from food contact surfaces using the methods disclosed
herein include bacterial and fungal species.
[0005] These and other aspects, objects, features, and advantages
of the example embodiments will become apparent to those having
ordinary skill in the art upon consideration of the following
detailed description of illustrated example embodiments.
BRIEF DESCRIPTION OF THE DRAWINGS
[0006] FIG. 1 is a graph showing Listeria monocytogenes populations
recovered on modified oxford medium from stainless steel coupons
post treatment with an example antimicrobial composition disclosed
herein. (4.4 mL spray nozzle)
[0007] FIG. 2 is a graph showing reduction in L. monocytogenes
populations on stainless steel coupons due to treatment with an
example antimicrobial composition disclosed herein.
[0008] FIG. 3 is a graph showing reduction in L. monocytogenes
populations recovered on modified oxford medium from stainless
steel coupons post-treatment with an example antimicrobial
composition disclosed herein (8 mL spray nozzle).
[0009] FIG. 4 is a graph showing reduction in L. monocytogenes
populations on stainless steel coupons due to treatment with an
example antimicrobial composition disclosed herein (8 mL spray
nozzle).
[0010] FIG. 5 is a graph showing L. monocytogenes populations
recovered on modified oxford medium from stainless steel coupons
(n=5) at various time points after treatment with an example
antimicrobial composition disclosed herein.
[0011] FIG. 6 is a graph showing reduction in L. monocytogenes
population on stainless steel coupons (n=5) at various time points
after treatment with an example antimicrobial composition disclosed
herein.
[0012] FIG. 7 is a graph showing L. monocytogenes population
recovered on modified oxford medium from stainless steel coupons
(n=5) at various time points after treatment with an example
antimicrobial composition disclosed herein.
[0013] FIG. 8 is a graph showing reduction in L. monocytogenes
population on stainless steel coupons (n=5) at various time points
after treatment with an example antimicrobial composition disclosed
herein.
DETAILED DESCRIPTION OF THE EXAMPLE EMBODIMENTS
Overview
[0014] Embodiments disclosed herein provide methods for
decontaminating food contact surfaces using a fine spray/mist that
comprise an antimicrobial composition comprising an organic acid
and an organic citrus extract. The spray application reduces the
microbiological populations on the surface of food contact
surfaces, including potential pathogens, thereby reducing
cross-contamination and improving food safety. The composition
described herein can be used to target a number of microbiological
pathogens, including L. monocytogenes, which is a known
cross-contaminant in retail environments such as deli counters and
slicers due to its ability to grow at refrigerated temperatures and
moist environments making it a known cause of post-processing
contamination of ready-to-eat food products. The composition is
applied to a surface to be treated for a period of time sufficient
to reduce or eliminate the microbial contamination.
[0015] In certain example embodiments, the antimicrobial
composition comprises an organic acid and an organic citrus
extract. The antimicrobial composition may also further comprise a
masker and an emulsifier. The antimicrobial composition may be used
to reduce or eliminate microbiological contamination on food
contact surfaces. Example food contact surfaces include, but are
not limited to, wood surfaces, glass surfaces, plastic surface,
rubber surfaces and stainless steel surfaces. In certain example
embodiments, the antimicrobial compositions achieve at least a 10%,
at least a 15%, at least a 20%, at least a 25%, at least a 30%, at
least 35%, at least 40%, at least 45%, at least 50%, at least 55%,
at least 60%, at least 65%, at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 97%, at
least 98%, or at least 99% reduction in microbial populations on
the food contact surface treated.
[0016] The composition may be applied as a fine spay or mist on the
surface to be treated. In certain example embodiments, the spray or
mist is applied at a rate of 1 mL/10 seconds of spray, 2 mL/10
seconds of spray, 3 mL/10 seconds of spray, 4 mL/10 seconds of
spray, 5 mL/10 seconds of spray, 6 mL/10 seconds of spray, 7 mL/10
seconds of spray, 8 mL/10 seconds of spray, 9 mL/10 seconds of
spray, 10 mL/10 seconds of spray, 11 mL/10 seconds of spray, 12
mL/10 seconds of spray, 13 mL/10 seconds of spray, 14 mL/10 seconds
of spray, 15 mL/10 seconds of spray, 16 mL/10 seconds of spray, 17
mL/10 seconds of spray, 18 mL/10 seconds of spray, 19 mL/10 seconds
of spray, 20 mL/10 seconds of spray, 21 mL/10 seconds of spray, or
22 mL/10 seconds of spray. The spray may be applied using a 4.4 mL,
8 mL, or 22 mL nozzle. The amount of spray applied is based on the
surface area to be treated. In certain example embodiments, an
amount of spray applied is sufficient to form a thin film on the
surface to be treated. The amount of time a spray should be
dispensed in order to apply a thin film to a surface to be treated
can be determined based on the type of spray nozzle used.
[0017] In certain example embodiments, the antimicrobial
composition is allowed to remain in contact with the treated
surface for at least 3 hours, at least 2 hours, at least 60 mins,
at least 55 mins, at least 50 mins, at least 45 mins, at least 40
mins, at least 35 mins, at least 30 mins, at leas 25 mins, at least
20 mins, at least 15 mins, at least 10 mins, at least 5 min, at
least 3 min, at least 2 min, at least 1 min, at least 30 second, at
least 20 second, at least 10 seconds, at least 5 seconds, or at
least 1 second before food is brought in contact with the treated
surface.
[0018] The organic acid may include acetic acid, formic acid,
propionic acid, butyric acid, valeric acid, caprioc acid, oxalic
acid, lactic acid, malic acid, citric acid, benzoic acid, carbonic
acid, or a combination thereof. In certain example embodiments, the
organic acid is acetic acid and/or a consumable salt equivalent
thereof In certain example embodiments, the acetic acid is sourced
from a vinegar. Source vinegar materials may include, for example,
corn, sugar cane, glacial acetic, and apple cider. Source vinegar
materials may include, for example, corn, sugar cane, glacial
acetic, and apple cider. The vinegar may be prepared using standard
buffering agents known in the art. In certain example embodiments,
the vinegar comprises 0.5% to 8% acetic acid. In certain other
example embodiments, the vinegar comprises 1% to 4% acetic acid. In
certain example embodiments, the source vinegar is a buffered
vinegar with an acidity of approximately 1.2% and pH of 6.9.
[0019] The organic acid may be at a concentration of 5% to 99.5%,
10% to 99.5%, 15% to 99.5%, 20% to 99.5%, 25% to 99.5%, 30% to
99.5%, 35% to 99.5%, 40% to 99.5%, 45% to 99.5%, 50% to 99.5%, 55%
to 99.5%, 60% to 99.5%, 65% to 99.5%, 70% to 99.5%, 75% to 99.5%,
80% to 99.5%, 85% to 99.5%, 90% to 99.5%, 91% to 99.5%, 92% to
99.5%, 93% to 99.5%, 94% to 99.5%, 95% to 99.5%, 96% to 99.5%, 97%
to 99.5%, or 98% to 99.5%, 5% to 50%, 10% to 50%, 15% to 50%, 20%
to 50%, 25% to 50%, 30% to 50%, 35% to 50%, 40% to 50%, 45% to 50%,
50% to 75%, 55% to 75%, 60% to 75%, 65% to 75%, or 70% to 75%
(w/v). In one example embodiment, the organic acid is at a
concentration between 95% and 99.5% (w/v).
[0020] In certain example embodiments, the organic acid has a pH of
approximately 5.5 to approximately 7.3, approximately 5.6 to
approximately 7.3, approximately 5.7 to approximately 7.3,
approximately 5.8 to approximately 7.3, approximately 5.9 to
approximately 7.3, approximately 6.0 to approximately 7.3,
approximately 6.1 to approximately 7.3, approximately 6.2 to
approximately 7.3, approximately 6.3 to approximately 7.3,
approximately 6.4 to approximately 7.3, approximately 6.5 to
approximately 7.3, approximately 6.6 to approximately 7.3,
approximately 6.7 to approximately 7.3, approximately 6.8 to
approximately 7.3, approximately 6.9 to approximately 7.3,
approximately 7.0 to approximately 7.3, approximately 7.1 to
approximately 7.3, approximately 7.2 to approximately 7.3,
approximately 5.5 to approximately 7.1, approximately 5.5 to
approximately 5.9, approximately 5.5 to approximately 5.8,
approximately 5.5 to approximately 5.7, approximately 5.5 to
approximately 5.6. As used in the context of describing pH value
ranges above, "approximately" means a pH value within 0.05 of the
stated pH values.
[0021] The organic citrus extract is derived from citrus fruits,
including but not limited to amanatsu, balady citron, bergamot
orange, bitter orange, blood orange, Buddha's hand, calamondin, cam
sanh, citron, clementine, Corsican citron, desert lime, etrog,
finger lime, Florentine citron, grapefruit, Greek citron,
hyuganatsu, iyokan, kabosu, kaffir lime, key lime, kinnow, kiyomi,
kumquat, lemon, lime, mandarin orange, mangshanyegan, Meyer lemon,
Moroccan citron, myrtle-leaved orange, orange, oroblanco, Persian
lime, pomelo, ponderosa lemon, rangpur, round lime, satsuma,
shangjuan, shonan gold, sudachi, sweet limetta, Taiwan tangerine,
tangelo, tangerine, tangor, ugli fruit, yuzu, or combination
thereof. In certain example embodiments, the citrus extract is
prepared from the albedo and flavedo layers of the citrus fruits.
In certain example embodiments, the citrus extract is prepared from
Citrus aurantium amara fruit extract, Citrus reticulate fruit
extract, and Citrus aurantium sinesis peel extract. The extract may
be suspended in glycerin from a natural source or other suitable
carrier. In certain example embodiments, the glycerin is a
vegetable glycerin. In certain example embodiments, the organic
citrus extract is a blend of two or more of the citrus fruits
listed above. In certain example embodiments, the organic citrus
extract is a blend of two or more citrus fruits, but not including
grapefruit derived extracts. In certain example embodiments, the
composition may have a honey color. In certain example embodiments,
the composition may be at least 70%, at least 75%, at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, or 100% soluble in water. In certain
example embodiments, the composition may be a crystalline liquid.
In certain example embodiments, the organic citrus fruit extract
comprises citrus bioflavonoids and citrus polyphenols at .gtoreq.4%
concentration and are an active ingredients in the formulation. In
certain example embodiments, the organic citrus extract is derived
from certified organic citrus fruits. In certain example
embodiments, the carrier is from a certified organic plant
material.
[0022] In certain example embodiments, the organic citrus extract
is at a concentration of 0.1% to 5%, 0.1% to 4.5%, 0.1% to 4.0%,
0.1% to 3.5%, 0.1% to 3.0%, 0.1% to 2.5%, 0.1% to 2.0%, 0.1% to
1.5%, 0.1% to 1.0%, 0.1% to 0.9%, 0.1% to 0.8%, 0.1% to 0.7%, 0.1%
to 0.6%, 0.1% to 0.6%, 0.1% to 0.5%, 0.1% to 0.4%, 0.1% to 0.3%,
0.11% to 0.2%, 0.2% to 1%, 0.3% to 1%, 0.4% to 1%, 0.5% to 1%, 0.6%
to 1%, 0.7% to 1%, 0.8% to 1%, 0.9% to 1%, 1% to 5%, 1.5% to 5%, 2%
to 5%, 2.5% to 5%, 3% to 5%, 3.5% to 4%, or 4.5% to 5% (w/v).
[0023] In certain example embodiments, the organic citrus extract
has a pH of approximately 2 to approximately 3.5, approximately 2
to approximately 3.4, approximately 2 to approximately 3.3,
approximately 2 to approximately 3.2, approximately 2 to
approximately 3.1, approximately 2 to approximately 3,
approximately 2 to approximately 2.9, approximately 2 to
approximately 2.8, approximately 2 to approximately 2.7,
approximately 2 to approximately 2.6, approximately 2 to
approximately 2.5, approximately 2 to approximately 2.4,
approximately 2 to approximately 2.3, approximately 2 to
approximately 2.2, approximately 2 to approximately 2.1,
approximately 2.1 to approximately 3.5, approximately 2.2 to
approximately 3.5, approximately 2.3 to approximately 3.5,
approximately 2.4 to approximately 3.5, approximately 2.5 to
approximately 3.5, approximately 2.6 to approximately 3.5,
approximately 2.7 to approximately 3.5, approximately 2.8 to
approximately 3.5, approximately 2.9 to approximately 3.5,
approximately 3 to approximately 3.5, approximately 3.1 to
approximately 3.5, approximately 3.2 to approximately 3.5,
approximately 3.3 to approximately 3.5, or approximately 3.4 to
approximately 3.5.
[0024] The antimicrobial composition may optionally further
comprise a masker. Any commercially available masker suitable for
suppressing bitterness and acidity may be used. In one example
embodiment the masker is a naturally derived masker. In certain
example embodiments, the masker may be at a concentration of
approximately 0.05% to 0.2%, 0.05% to 0.1%, 0.05% to 0.09%, 0.05%
to 0.08%, 0.05% to 0.07%, or 0.05% to 0.06% w/v.
[0025] The antimicrobial composition may optionally also comprise
an emulsifier, used as a processing aid. Any commercially available
emulsifier suitable for allowing stable liquid suspension may be
used. In certain example embodiments, the emulsifier may be at a
concentration of approximately 0.1% to 0.15%, 0.1% to 0.2%, 0.5% to
1%, 1% to 1.5, 1.5% to 1.9% or at levels .ltoreq.2% w/v.
[0026] The antimicrobial compositions described herein may be used
to reduce microbial contamination on food contact surfaces from one
or more microbes. The microbe may be a pathogenic microbe. In
certain example embodiments, the microbe is a bacteria. The
bacteria may be a psychrotroph, a coliform, a lactic acid bacteria,
or a spore-forming bacteria. In certain example embodiments, the
bacteria is a Staphylococcus species, a Pseudomonas species, a
Micrococcus species, an Aerococcus species, a Lactococcus species,
a Leuconostoc species, a Streptococcus species, a Bacillus species,
a Clostridium species, a Eubacteria species, an Enterococcus
species, a Listeria species, or a combination thereof. In certain
example embodiments, the bacteria is a Listeria species. In certain
example embodiments, the Listeria species is L. monocytogenes.
[0027] In certain example embodiments the microbe a yeast. In
certain example embodiments, the fungi is a yeast. Example yeast
may include, but are not limited to, a Geotrichum species, a
Saccharomyces species, a Hansenula species, a Candida species, a
Kluyveromyces species, a Debaryomyces species, a Pichia species, or
combination thereof
[0028] The invention is further described in the following
examples, which do not limit the scope of the invention described
in the claims.
EXAMPLES
Example 1
Bacterial Cultures and Inoculum Preparation:
[0029] Bacterial Cultures were obtained from American Type Culture
Collection (ATCC). A five-strain (ATCC 19112, 19118, 19115, 13932,
and 19111) cocktail of Listeria monocytogenes was used for
inoculation of coupon. For inoculum preparation, freeze-dried
cultures were grown in Tryptic Soy Broth (TSB) and fresh overnight
24 hr grown cultures were used. Each strain was combined in to a
single mixed culture suspension to obtain a five-strain cocktail.
The cell density of the inoculum was determined by spread plating
and incubating the plates at 35.degree. C. for 24 hr. The target
inoculum level for this study was 6-7 log CFU/coupon.
Sample Preparation:
[0030] Polished stainless steel coupons were cleaned and autoclaved
for use. Binder clips were attached to the coupons to allow hanging
them from a small rod.
Inoculation of Samples:
[0031] Sterile stainless steel coupons were dipped in a 24 hr grown
L. monocytogenes five-strain cocktail for 30 seconds and then hung
to dry in a biosafety cabinet to allow bacterial attachment for
30-40 min. Excess inoculum on the inoculum was allowed to drip
down.
Treatment of Samples:
[0032] Samples were treated with the antimicrobial solution using
the spray/mist method using aerosolized cans under a biosafety
cabinet. The antimicrobial was applied in different ways to assess
its effect on the bacterium attached to the stainless steel coupon
surface.
[0033] Treatment Formulation: [0034] Vinegar [0035] Organic Citrus
extract [0036] Masker
[0037] Emulsifier
[0038] Spray Cans Used: [0039] 4.4 ml nozzle (dispenses 4.4 ml of
antimicrobial when sprayed for 10 s) [0040] 8 ml nozzle (dispenses
8 ml of antimicrobial when sprayed for 10 s)
Bacterial Enumeration:
[0041] After treatment application, each stainless steel coupon was
individually placed in a 50 ml sterile conical tube with 15 ml of
0.1% sterile peptone diluent. Bacterial cells attached on the
surface of coupon were dislodged by vortexing for 1 min. Serial
dilutions were then prepared and spread plated on to selective
media for Listeria monocytogenes, modified Oxford Medium (MOX). The
plates were incubated at 35.degree. C. for 48 h.
Test 1: 4.4 ml Nozzle and Individually Treated Coupons
[0042] Sterile stainless steel coupons were surface inoculated with
bacterial cocktail. 4.4 ml spray nozzle was used for antimicrobial
composition application. Each coupon was individually sprayed with
the antimicrobial composition for 10 secs on both sides, amounting
to a 4.4 ml total antimicrobial composition on the surface of each
coupon. The coupons were then held for 30 sec, 1 min, 2 min, 3 min
and 5 min to allow for exposure to the antimicrobial composition.
After the specified contact time, bacterial enumeration on the
surface post treatment was performed. Control samples received no
treatment and indicated initial levels of bacterial populations
attached to the surface of the stainless steel coupons. Total
number of samples analyzed were as follows: Control (n=16), 30 s
(n=9), 1 min (n=9), 2 min (n=9), 3 min (n=14), 5 min (n=9). The
data was generated from 3 separate days of testing and samples were
added together to obtain an average for each subset. The results
presented are based on an average of each treatment subset. Highest
bacterial reductions were seen with 1 min antimicrobial contact
time. See FIGS. 1 and 2.
Test 2: 8 ml Nozzle and Individually Treated Coupons
[0043] Sterile stainless steel coupons were surface inoculated with
bacterial cocktail. 8 ml spray nozzle was used for antimicrobial
application. Each coupon was individually sprayed with the
antimicrobial for 10 sec on both sides, amounting to 8 ml total
antimicrobial composition on the surface of each coupon. The
coupons were then held for 2 min, 3 min and 5 min to allow exposure
to antimicrobial. After specified contact time, bacterial
enumeration on the surface post treatment was performed. Control
samples received no treatment and indicated initial levels of
bacterial populations attached to the surface of the stainless
steel coupons. Total number of samples analyzed were as follows:
Control (n=3), 2 min (n=4), 3 min (n=4), 5 min (n=4). The results
presented are based on average of each treatment subset. Bacterial
reductions increased with increased contact time with the
antimicrobial composition. See FIGS. 3 and 4.
Test 3: 8 ml Nozzle and Bulk Sprayed Coupons
[0044] Testing was continued to determine the effect of a
non-specific dosage application of the antimicrobial treatment on
the stainless steel coupon surface. This was performed to mimic
food processing surface area conditions where the area to which the
treatment applied may vary and precise application based on surface
area may not be possible. Thirty sterile stainless steel coupons
were surface inoculated with bacterial cocktail. 8 ml spray nozzle
was used for antimicrobial composition application. The coupons
were then divided in to 6 subsets and treated as below: [0045]
T1-Control (n=5, no treatment) Samples received no treatment and
indicated initial levels of bacterial populations attached to the
surface of the stainless steel coupons. [0046] T2-Bulkspray 8 ml
for 10 s All coupons were sprayed together using spray can with 8
ml spray nozzle [0047] T3-Reinoculation In order to mimic
re-contamination scenario in a food processing area, e.g. delis,
samples were re-inoculated with low level (approx. 4 log cfu/ml) of
five-strain L. monocytogenes cocktail by dipping coupons for 15 s
in the cocktail). [0048] T4-Bulk spray 8 ml for 10 s. All coupons
were sprayed together using spray can with 8 ml spray nozzle.
[0049] T5-Reinoculation was repeated as performed in T3 [0050]
T6-Bulk spray 8 ml for 10 s Same as T2 and T4
[0051] The results presented are based on average (n=5) of each
treatment subset. See FIGS. 5 and 6.
Test 4: 8 mL Nozzle and Bulk Sprayed Coupons
[0052] This was a modification of test 3 to evaluate if the initial
bacterial load could be reduced even further by increasing the
amount of antimicrobial composition applied at treatment 2 stage.
In test 4, treatment 2, an 8 ml nozzle was used to spray all the
coupons together for 20 sec.
[0053] Thirty sterile stainless steel coupons were surface
inoculated with bacterial cocktail. 8 ml spray nozzle was used for
antimicrobial application. The coupons were then divided in to 6
subsets and treated as below: [0054] T1-Control (n=5, no treatment)
Samples received no treatment and indicated initial levels of
bacterial populations attached to the surface of the stainless
steel coupons. [0055] T2-Bulkspray 8 ml for 20 s All coupons were
sprayed together using spray can with 8 ml spray nozzle [0056]
T3-Reinoculation In order to mimic re-contamination scenario in a
food processing area, e.g. delis, samples were re-inoculated with
low level (approx. 4 log cfu/ml) of five-strain L. monocytogenes
cocktail by dipping coupons for 15 s in the cocktail) [0057]
T4-Bulk spray 8 ml for 10 sec. All coupons were sprayed together
using spray can with 8 ml spray nozzle [0058]
T5-Reinoculation-Reinoculation was repeated as performed in T3
[0059] T6-Bulk spray 8 ml for 10 s--same as T2 and T4
[0060] The results presented are based on average (n=5) of each
treatment subset. See FIGS. 7 and 8.
Test 5: 8 ml Nozzle and Bulk Sprayed Coupons
[0061] This was a modification of test 3 and 4 to evaluate
effectiveness of antimicrobial spray on the surface by allowing
antimicrobial to dry on the coupon surface before re-introduction
of the pathogen on the surface and then allowing it to attach again
for 30 min before re-applying treatment. This test would mimic food
processing and handling conditions in a regular ongoing work shift
where decontamination and re-contamination happens throughout the
processing time period.
[0062] Thirty sterile stainless steel coupons were surface
inoculated with bacterial cocktail. 8 ml spray nozzle was used for
antimicrobial application. The coupons were then divided in to 6
subsets and treated as below: [0063] T1-Control (n=5, no
treatment)--Samples received no treatment and indicated initial
levels of bacterial populations attached to the surface of the
stainless steel coupons. [0064] T2-Bulk spray 8 ml for 20 sec--All
coupons were sprayed together using spray can with 8 ml spray
nozzle and allowed to dry for 1 h with antimicrobial spray on the
surface. [0065] T3-In order to mimic re-contamination scenario in a
food processing area, e.g., delis, samples were re-inoculated with
low level (approx. 4 log cfu/ml) of five-strain L. monocytogenes
cocktail by dipping coupons for 15 s in the cocktail). The coupons
were allowed to dry for 30 min to allow re-attachment of bacterial
cells. [0066] T4-Bulk spray 8 ml for 10 s--All coupons were sprayed
together using spray can with 8 ml spray nozzle and allowed to dry
for 1 h with antimicrobial on the surface [0067] T5-Reinoculation
was repeated as performed in T3 [0068] T6-Bulk spray 8 ml for 10
sec--Same as T2 and T4
[0069] The results presented are based on average (n=5) of each
treatment subset. The results indicate that bacterial cells are
unable to attach the surface in the presence of an example
antimicrobial compositions disclosed herein. Bacterial log
reductions continue to increase as more antimicrobial composition
is applied on the surface in spite of re-contamination.
Example 2
Food Contact Surface Testing
Stainless Steel Surface
[0070] A five strain cocktail of Listeria monocytogenes was used to
inoculate a representative food contact stainless steel surface
(surface area 2419.5 cm.sup.2) using the sponge method. Sterile
sponge moistened with the five-strain cocktail suspension was used
to inoculate the surface. The bacterial cultures were allowed to
attach on the surface for 20 min. Spray treatment (8 ml/10 s) was
used to treat the surface. Following treatment was used:
[0071] Treatment-20 s spray, 1 h hold time
[0072] Control-Non treatment
[0073] Sponge samples were then taken to enumerate bacterial
population on the surface before and after treatments. Modified
oxford medium (MOX) was used and plates were incubated at
35.degree. C. for 24 hours. The results indicate reduction in
bacterial populations due to treatment.
TABLE-US-00001 TABLE 1 Sample Bacterial Count Recovered Bacterial
Reduction Control 21 CFU/cm.sup.2 (No Treatment) Treatment 0.62
CFU/cm.sup.2 20.38 CFU/cm.sup.2 (1 hr hold time)
Treatment of HDPE Cutting Board
[0074] A five strain cocktail of Listeria monocytogenes was used to
inoculate two cutting boards, (surface areas 644.35 cm.sup.2 and
769.35 cm.sup.2) using the sponge method. Sterile sponge moistened
with the five-strain cocktail suspension was used to inoculate the
surface. The bacterial cultures were allowed to attach on the
surface for 20 min. Spray treatment (8 ml/10 s) was used to treat
these surfaces. Sponge samples were taken before and after
treatment from the cutting boards. Bacteria were enumerated on
modified oxford medium (MOX)
TABLE-US-00002 TABLE 2 Cutting Board Area 769 cm.sup.2 Bacterial
Count Recovered Bacterial Reduction Control 2.94 Log CFU/cm (No
Treatment) Treatment 1.72 Log CFU/cm 1.22 Log CFU/cm.sup.2 (1 hr
hold time)
TABLE-US-00003 TABLE 3 Cutting Board Area 769 cm.sup.2 Bacterial
Count Recovered Bacterial Reduction Control 4.67 Log CFU/cm.sup.2
(No Treatment) Treatment 2.69 Log CFU/cm.sup.2 1.98 CFU/cm2 (10 sec
spray with 8 ml nozzle)
TABLE-US-00004 TABLE 4 Cutting Board Area 644.35 cm.sup.2 Bacterial
Count Recovered Bacterial Reduction Control 4.43 Log CFU/cm.sup.2
(No Treatment) Treatment 3.07 Log CFU/cm.sup.2 1.36 Log CFU/cm2 (10
s spray with 8 ml nozzle, 3 min hold time)
Total Aerobic Count
[0075] Cutting boards (surface areas 644.35 cm.sup.2 and 769.35
cm.sup.2) were inoculated with total aerobic bacteria using sponge
method. Bacterial cultures were allowed to attach for 20 min. Spray
treatment (8 ml/10 s) was applied for 10 sec and held for 3 min and
1 hr. Bacterial reductions were calculated due to treatment by
comparing bacterial populations recovered before and after
treatment. Bacteria were enumerated on tryptic soy agar (TSA).
TABLE-US-00005 TABLE 5 Cutting Board Area 769 cm.sup.2 Bacterial
Count Recovered Bacterial Reduction Control 4.12 Log CFU/cm.sup.2
(No Treatment) Treatment 3.26 Log CFU/cm.sup.2 0.86 Log CFU/cm2 (10
s spray with 8 ml nozzle, 1 hr hold time)
TABLE-US-00006 TABLE 6 Cutting Board Area 644.35 cm.sup.2 Bacterial
Count Recovered Bacterial Reduction Control 4.46 Log CFU/cm.sup.2
(No Treatment) Treatment 3.41 Log CFU/cm.sup.2 1.05 Log CFU/cm2 (10
s spray with 8 ml nozzle, 3 min hold time)
[0076] Various modifications and variations of the described
methods and compositions of the disclosure will be apparent to
those skilled in the art without departing from the scope and
spirit of the invention. Although the invention has been described
in connection with specific embodiments, it will be understood that
it is capable of further modifications and that the invention as
claimed should not be unduly limited to such specific embodiments.
Indeed, various modifications of the described modes for carrying
out the invention that are obvious to those skilled in the art are
intended to be within the scope of the invention. This application
is intended to cover any variations, uses, or adaptations of the
invention following, in general, the principles of the invention
and including such departures from the present disclosure come
within known customary practice within the art to which the
invention pertains and may be applied to the essential features
herein before set forth.
[0077] All publications, patents, and patent applications mentioned
herein are incorporated by reference to the same extent as if each
individual publication, patent or patent application was
specifically and individually indicated to be incorporated by
reference in its entirety. In the event of there being a difference
between definitions set forth in this application and those in
documents incorporated herein by reference, the definitions set
forth herein control.
* * * * *